Method for the generation of monoclonal antibodies derived from human b cells

a monoclonal antibody and human b cell technology, applied in the field of human b cells, can solve the problems of low cloning efficiency, elusive efficient method of obtaining such antibodies, and low efficiency of immortalization

Inactive Publication Date: 2013-01-31
DUKE UNIV
View PDF0 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

An efficient method of obtaining such antibodies has, however, been elusive.
The efficiency of immortalization, however, can be low, as can the cloning efficiency of the immortalized B cells.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Method for the generation of monoclonal antibodies derived from human b cells
  • Method for the generation of monoclonal antibodies derived from human b cells
  • Method for the generation of monoclonal antibodies derived from human b cells

Examples

Experimental program
Comparison scheme
Effect test

example

Experimental Details

Preparation of Complete Medium

[0029]Complete Medium (CM) for PBMC, EBV-B cells, J774A.1, and K6H6 / B5 cell lines

[0030]RPMI 1640 (Invitrogen) supplemented with:[0031]1) 15.2% heat-inactivated fetal calf serum (FCS)[0032]2) 1% non-essential amino acids (NEAA)[0033]3) 1 mM sodium pyruvate[0034]4) 15 mM HEPES buffer, pH 7.3[0035]5) 2 mM L-glutamine (Glu)[0036]6) 100 U / ml penicillin G (Pen)[0037]7) 100 μg / ml streptomycin (Strep)

[0038]Preparation:[0039]1. Thaw a bottle of FCS (500 ml / bottle, −20° C.) and incubate at 56° C. for 30 min for heat inactivation (inactivation of c′)[0040]2. Let bottle cool down and add 31 ml of Glu+Pen / Strep (100×, 100 ml / bottle, −20° C.)[0041]3. Aliquot the FCS-antibiotics mixture into tubes at 50 ml / tube and freeze at −20° C.[0042]4. Add 100 ml of FCS-antibiotics mixture to bottle of RPMI 1640 (500 ml / bottle, 4° C.)[0043]5. Add 6.2 ml of HEPES buffer (100×, 100 ml / bottle, 4° C.) into the bottle[0044]6. Add 6.2 ml of sodium pyruvate (100×, 1...

experiment 8-28

[0136]

FrequencySampleof positiveCDR3IDPCR wellsVDHJMutatedlengthProductive113 / 20 1~69*023~3*014*020.0372499F211 / 20 4~39*066~13*011*010.04301015F1 / 203~49*032~2*01 / inv, 02 / inv4*020.0925372N34 / 203~23*012~2*01 / inv, 02 / inv3*020.02949117F43 / 204~39*012~21*025*010.05221919F53 / 202~5*104~17*014*020.04143613F613 / 20 4~31*030~IR*01C4*020.02193810F73 / 204~34*012~2*01, 024*020.00266718F88 / 204~59*012~2*024*010.03753418FTubeOriginalSortCDR3IgIDWellWellVDHJMutatedlengthIsotypeProductive1B7A101~69*023~3*014*020.0374639779G1F1B7A101~69*023~3*014*020.0372492849G1F1B7A101~69*023~3*014*020.0373563229G1F1B7A21~69*023~3*014*020.0401146139G1F1B7A21~69*023~3*014*020.0401146139G1F1B7A21~69*023~3*014*020.0402298859G1F1B7A31~69*022~2*01, 021*010.0744985679MN1B7A31~69*023~3*014*020.0372492849G1F1B7A31~69*023~10*01 / inv1*010.0442477889A1N1B7A41~69*023~3*014*020.0372492849G1F1B7A41~69*023~3*014*020.0372492849G1F1B7A41~69*023~3*014*020.0373563229G1F1B7A51~69*023~3*014*020.0429799439G1F1B7A51~69*023~3*014*020.037249284...

experiment 8-29

Experiment 8-30—Total IgG ELISA

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

PropertyMeasurementUnit
densityaaaaaaaaaa
cell densitiesaaaaaaaaaa
fluorescentaaaaaaaaaa
Login to view more

Abstract

The present invention relates, in general, to human B cells, and, in particular to a method of immortalizing and cloning human B cells and to monoclonal antibodies derived therefrom. The invention further relates to methods of using the monoclonal antibodies for therapeutic and diagnostic purposes.

Description

[0001]This application claims priority from U.S. Provisional Application No. 61 / 322,725, filed Apr. 9, 2010 and U.S. Provisional Application No. 61 / 322,821 filed Apr. 10, 2010, the entire contents of which are incorporated herein by reference.[0002]This invention was made with government support under Grant No. AI067854-02 awarded by the National Institutes of Health. The government has certain rights in the invention.TECHNICAL FIELD[0003]The present invention relates, in general, to human B cells, and, in particular to a method of immortalizing and cloning human B cells and to monoclonal antibodies derived therefrom. The invention further relates to methods of using the monoclonal antibodies for therapeutic and diagnostic purposes.BACKGROUND[0004]The isolation and characterization of monoclonal antibodies that neutralize, for example, a broad spectrum of HIV-1 isolates are important to the design of an effective HIV-1 vaccine. An efficient method of obtaining such antibodies has, h...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Applications(United States)
IPC IPC(8): C12N15/86
CPCC07K16/00C07K2317/76C07K16/1045
Inventor HAYNES, BARTON F.BONSIGNORI, MATTIAHWANG, KWAN-KILIAO, HUA-XINMONTEFIORI, DAVID C.LUFTIG, MICAH A.
Owner DUKE UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap